Application of dithio metal ligand compound in preparation of antitumor drugs
By developing dithio metal ligand compounds, the problem of high toxic and side effects of existing platinum anti-cancer drugs has been solved, effective inhibition of a variety of tumor cells has been achieved, and broad-spectrum anti-tumor activity and low-cost characteristics are achieved.
Patent Information
- Application Number
- CN202410178602.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-02-09
- Publication Date
- 2025-08-12
AI Technical Summary
Existing platinum anti-cancer drugs have problems with high toxic side effects and high cost, and other metal drugs are still rare in clinical applications, making it difficult to find anti-cancer drugs with broad-spectrum and small toxic side effects.
A dithio metal ligand compound with strong redox activity was developed to prepare anti-tumor drugs. By dropping the metal salt solution to the dithio ligand solution to form a precipitate, the compound was obtained after treatment, and its structure was determined by electrospray mass spectrometry and single crystal structure analysis.
Dithio metal ligand compounds show good inhibitory ability on lung, liver and gastric cancer cells, have broad-spectrum anti-tumor activity, have small toxic side effects, low cost, and have wide application prospects.
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Abstract
Description
Technical Field
[0001] The present invention relates to the field of metal ligand compounds, in particular to the application of disulfide metal ligand compounds in the preparation of anti-tumor drugs. Background Art
[0002] Cancer is a common and frequently occurring disease that poses a serious threat to human life. Its mortality rate ranks second only to cardiovascular disease, and it is considered another major cause of death. Metal complexes offer numerous unique advantages over other types of drugs. Different metal centers have varying coordination numbers and geometric configurations, facilitating the realization of diverse redox states. The thermodynamic and kinetic properties of metal complexes, as well as the unique properties of metal ions and ligands, offer unlimited potential for the application of metal complexes in the field of anti-tumor drugs.
[0003] Cisplatin and other approved second-generation drugs are commonly used metal complexes in cancer chemotherapy. However, platinum-based anticancer drugs are associated with numerous toxic side effects, affecting multiple tissues and organs throughout the body. Common clinical manifestations include neurotoxicity, hepatotoxicity, nephrotoxicity, ototoxicity, and myelotoxicity. These toxic effects are common and occur frequently, leading to treatment interruption due to patients' intolerance to these severe side effects. Long-term use can also lead to drug resistance, rendering treatment ineffective. This limits the clinical application of platinum-based anticancer drugs. Platinum, as a precious metal, contributes to the high cost of platinum-based chemotherapy drugs. To overcome these limitations of cisplatin-based drugs and to develop broader-spectrum, less toxic anticancer drugs, the design, synthesis, and application of novel metal anticancer drugs have become a focus of research. While several other metal complexes have been shown to exhibit antitumor activity, such as Au(I,III), Fe(II), Pt(II), Ru(II,III), and V(IV), few other metal-based drugs have been clinically applied. Summary of the Invention
[0004] In view of the shortcomings of the existing technology, the present invention provides a new disulfide metal ligand compound with redox activity, and for the first time uses it as a broad-spectrum anticancer drug, and finds that it has good tumor cell inhibitory activity.
[0005] To achieve this object, the present invention adopts the following technical solutions:
[0006] The present invention provides an application of a disulfide metal ligand compound in the preparation of an anti-tumor drug. The chemical structure of the compound is shown in formula (I):
[0007]
[0008] wherein R1 and R2 are each independently selected from any one of a hydrogen atom, an alkyl group, a substituted alkyl group, an alkenyl group, a substituted alkenyl group, an alkynyl group, a substituted alkynyl group, a monocyclic aromatic group, a substituted monocyclic aromatic group, a polycyclic aromatic group, a substituted polycyclic aromatic group, a polyheterocyclic aromatic group, a substituted polyheterocyclic aromatic group, and a polymer group having a molecular weight of 100,000 g / mol or less and containing a repeating unit structure of polyvinyl alcohol or polyethylene glycol;
[0009] A is selected from C, N or O;
[0010] M is a metal atom selected from any one of Fe, Co, Ni, Ru, Rh, Pd, Os, Ir, Pt, Mn, Tc, Re, Cr, Mo, W, V, Nb, Ta, Ti, Zr, Hf, Sc, Y, La, Ac, Cu, Zn, Ag, Cd, Au, Hg, Rg, Cn, Hs, Mt, Ds, Bh, Sg, Db, and Rf;
[0011] n is an integer greater than or equal to 2.
[0012] In some embodiments of the present invention, R1 and R2 are each independently selected from hydrogen, a C1-C4 alkyl group, and a monocyclic aromatic group; A is selected from N, O, and C; M is selected from Fe, Co, Ni, and Cu; and n is 2 or 3.
[0013] In some embodiments of the present invention, the chemical structure of the compound is shown in Formula (II) or Formula (III):
[0014]
[0015] In some embodiments of the present invention, the chemical structure of the compound is selected from the following group:
[0016]
[0017]
[0018] The present invention provides a method for preparing the disulfide metal ligand compound, comprising the following steps: dropwise adding a metal salt solution into a disulfide ligand solution to generate a precipitate, and post-treating the solution to obtain the disulfide metal ligand compound.
[0019] In some embodiments of the present invention, the molar ratio of the metal ion to the disulfide ligand is n:1.
[0020] In some embodiments of the present invention, the reaction temperature is room temperature; the reaction time is more than 6 hours, and can be 6-12 hours, 12-18 hours, 18-24 hours or longer, specifically 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 15 hours, 20 hours, 25 hours or longer.
[0021] In some embodiments of the present invention, the post-treatment comprises filtering, washing and vacuum drying the precipitate.
[0022] In some embodiments of the present invention, the filtration is a Buchner funnel filtration.
[0023] In some embodiments of the present invention, the washing medium is selected from methanol, ethanol or ultrapure water.
[0024] There is no particular limitation on the number of washing times in the present invention. To ensure sufficient washing without causing excessive product loss, three times is preferred.
[0025] In some embodiments of the present invention, the vacuum drying temperature is 40-60°C, which can be 40-45°C, 45-50°C, 50-55°C or 55-60°C, specifically 40°C, 50°C, or 60°C, and the vacuum drying time is 6-24h, which can be 6-12h, 12-18h or 18-24h, specifically 6h, 7h, 8h, 9h, 10h, 12h, 14h, 16h, 18h, 20h, 22h, or 24h. In some preferred embodiments of the present invention, the vacuum drying temperature is 60°C and the vacuum drying time is 12h.
[0026] The present invention determines the structure of the disulfide metal ligand compound by electrospray mass spectrometry and single crystal structure analysis.
[0027] The present invention uses cell experiments to study the effects of the above-mentioned disulfide metal ligand compound on tumor cells. The results show that the above-mentioned disulfide metal ligand compound has good inhibitory ability against three human tumor cells: lung cancer A459, liver cancer HEPG2, and gastric cancer SGC790 cells, has broad-spectrum anti-tumor activity, and has good application prospects in the preparation of anti-tumor drugs.
[0028] In some embodiments of the present invention, the anti-tumor drug further comprises a pharmaceutically acceptable carrier or excipient, or a combination thereof. The anti-tumor drug can be administered via conventional routes, including (but not limited to): intravenous injection, intravenous drip, subcutaneous injection, local injection, intramuscular injection, intratumor injection, intraperitoneal injection (e.g., intraperitoneal injection), intracranial injection, intracavitary injection, inhalation administration, implantation, etc.
[0029] The term "pharmaceutically acceptable" as used herein means that the drugs will not produce adverse, allergic or other untoward reactions when appropriately administered to animals or humans.
[0030] "Pharmaceutically acceptable excipients" should be compatible with the active ingredient, meaning they can be blended with it without significantly reducing the drug's effectiveness under normal circumstances. Specific examples of substances that can serve as pharmaceutically acceptable excipients include sugars, starch, cellulose and its derivatives, tragacanth powder, malt, gelatin, talc, solid lubricants, calcium sulfate, vegetable oils, alcohols, alginic acid, emulsifiers, wetting agents, surfactants, lyoprotectants, colorants, flavorings, tableting agents, stabilizers, diluents, excipients, antioxidants, preservatives, pyrogen-free water, isotonic saline solutions, buffers, and combinations thereof. These substances are used, as needed, to improve the stability of the formulation, enhance the activity or bioavailability of the drug, or produce an acceptable taste or odor when administered orally.
[0031] The pharmaceutical composition of the present invention can be prepared into an inhalation atomized preparation (such as a dry powder preparation, an aerosol preparation, an inhalation mist droplet preparation, etc.), an implantable gel preparation, a microneedle preparation, and can also be prepared into an injection form, for example, using physiological saline or an aqueous solution containing glucose and other adjuvants by conventional methods.
[0032] Compared with the prior art, the present invention has the following beneficial effects:
[0033] The disulfide metal ligand compound provided by the present invention is simple to prepare, low in cost, has a broad-spectrum anticancer effect, has less toxic and side effects, and has good application prospects in the preparation of anti-tumor drugs. BRIEF DESCRIPTION OF THE DRAWINGS
[0034] Figure 1 This is the ESI-MS spectrum of the product obtained in Example 1 of the present invention.
[0035] Figure 2 This is a structural diagram of the Fe(DTC)3 crystal obtained in Example 1 of the present invention.
[0036] Figure 3 This is the ESI-MS spectrum of the product obtained in Example 2 of the present invention.
[0037] Figure 4 This is a structural diagram of the Co(DTC)3 crystal obtained in Example 2 of the present invention. DETAILED DESCRIPTION
[0038] Before further describing the specific embodiments of the present invention, it should be understood that the scope of the present invention is not limited to the specific embodiments described below. It should also be understood that the terminology used in the examples is intended to describe specific embodiments and is not intended to limit the scope of the present invention. The experimental procedures in the following examples, where specific conditions are not specified, were generally performed under conventional conditions or according to the conditions recommended by the respective manufacturers.
[0039] When the embodiments provide numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, according to the understanding of the prior art by those skilled in the art and the description of the present invention, any methods, equipment, and materials of the prior art similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention may also be used to implement the present invention.
[0040] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related fields in the art.
[0041] Example 1
[0042] 1.1 Synthesis of disulfide metal ligand compound Fe(DTC)3
[0043] The disulfide metal ligand compound Fe(DTC)3 was synthesized in this embodiment, and its chemical structure is shown in the following formula (II):
[0044]
[0045] The synthesis method is as follows:
[0046] Accurately weigh 2.62g (0.75mmol) of sodium diethyldithiocarbamate ((C2H5)2NCSSNa), dissolve it in 20mL of ultrapure water and place it in a 100mL eggplant-shaped bottle. Accurately weigh 675mg (0.25mmol) of iron (III) chloride hexahydrate (FeCl3·6H2O) and dissolve it in 20mL of ultrapure water. Add it dropwise to the eggplant-shaped bottle and stir the reaction at room temperature for more than 6 hours to obtain a black precipitate. After filtering with a Buchner funnel and washing it three times with ultrapure water, place it in a vacuum drying oven at 60°C and dry it for 12 hours to obtain a black solid. Its ESI-MS spectrum is shown below. Figure 1 As shown in the figure, the molecular ion peak 500.010 in positive ion mode is consistent with the theoretical value (500.01).
[0047] Fe(DTC)3 crystals were obtained by solvent evaporation method. The crystal structure is shown in the figure below. Figure 2 As shown, analysis shows that the complex molecule is composed of a metal ion Fe(1) coordinated by three DTC ligands, and Co(1) is in a six-coordinate environment coordinated with S, which is consistent with the theoretical structure.
[0048] 1.2 Antitumor activity of disulfide metal ligand Fe(DTC)3
[0049] The following uses three common tumor cells (lung cancer A459, liver cancer HEPG2, and gastric cancer SGC790 cells) as examples to verify the anti-tumor activity of the disulfide metal ligand compound Fe(DTC)3. The specific experimental steps are as follows:
[0050] S1, human tumor cells A459, HEPG2, and SGC790 were cultured in DMEM high-glucose medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C in a 5% CO2 incubator. When cells reached over 80% confluency, the medium was discarded and the cells were digested with 0.8% trypsin containing 0.02% EDTA at 37°C. The cells were centrifuged, the supernatant discarded, and fresh culture medium was added. The cells were subcultured at a 1:3 ratio. Well-grown cells in the logarithmic growth phase were used for experiments.
[0051] S2. The cells in the logarithmic growth phase were digested with trypsin and counted under a microscope to prepare 6×10 4 Take 100 μl of the cell suspension to 96-well culture plates, and inoculate two duplicate wells of each plate for each cell type as duplicate wells. 3 cells / well, 100 μl of culture medium was used as blank control, and the cells were cultured at 37°C overnight.
[0052] S3. Prepare a stock solution of Fe(DTC)3 complex with an initial concentration of 20 mM, dilute it according to a concentration gradient, and add three replicates of each concentration to a 96-well plate. After treating the cells for 72 hours, mix Cell Counting Kit-8 (CCK-8) and serum-free essential basic medium in a 1:10 volume ratio, add 100 μL per well to the wells to be tested, and incubate in a 37°C, 5% CO2 incubator for 1 hour.
[0053] S4. Absorbance at 450 nm was measured using a microplate reader, and the values for each plate were recorded. Cell viability in each group was determined using the CCK-8 assay, and the inhibition rate was calculated: inhibition rate = (OD value of the control group - OD value of the drug group) / (OD value of the control group) × 100%. Calculation of the half-maximal inhibitory concentration (IC50) values: GraphPad Prism 9 software was used to calculate the half-maximal inhibitory concentration (IC50) values based on the effects of different drug concentrations on cells, as shown in Table 1.
[0054] Example 2
[0055] 2.1 Synthesis of disulfide metal ligand compound Co(DTC)3
[0056] The disulfide metal ligand compound Co(DTC)3 was synthesized in this embodiment, and its chemical structure is shown in the following formula (III):
[0057]
[0058] The synthesis method is as follows:
[0059] Accurately weigh 2.62 g (0.75 mmol) of sodium diethyldithiocarbamate ((C2H5)2NCSSNa), dissolve it in 20 mL of ultrapure water and place it in a 100 mL eggplant-shaped bottle. Accurately weigh 680 mg (0.25 mmol) of hexaaminocobalt (III) chloride.
[0060] (Cl3CoH 18 N6) was dissolved in 20 mL of ultrapure water and added dropwise to the eggplant-shaped flask. The mixture was stirred at room temperature for more than 6 h to obtain a green precipitate. The precipitate was filtered through a Buchner funnel and washed three times with ultrapure water. The precipitate was then dried in a vacuum drying oven at 60°C for 12 h to obtain a green solid. The ESI-MS spectrum is shown below. Figure 3 As shown in the figure, the molecular ion peak 503.008 in positive ion mode is consistent with the theoretical value (503.01).
[0061] Co(DTC)3 crystals were obtained by solvent evaporation method. The crystal structure is shown in the figure below. Figure 4 As shown, analysis shows that the complex molecule is composed of a metal ion Co(1) and three DTC ligands, and Co(1) is in a six-coordinate environment coordinated with S, which is consistent with the theoretical structure.
[0062] 2.2 Antitumor activity of disulfide metal ligand compound Co(DTC)3
[0063] The following uses three common tumor cells (lung cancer A459, liver cancer HEPG2, and gastric cancer SGC790 cells) as examples to verify the anti-tumor activity of the disulfide metal ligand compound Co(DTC)3. The specific experimental steps are as follows:
[0064] S1, human tumor cells A459, HEPG2, and SGC790 were cultured in DMEM high-glucose medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C in a 5% CO2 incubator. When cells reached over 80% confluency, the medium was discarded and the cells were digested with 0.8% trypsin containing 0.02% EDTA at 37°C. The cells were centrifuged, the supernatant discarded, and fresh culture medium was added. The cells were subcultured at a 1:3 ratio. Well-grown cells in the logarithmic growth phase were used for experiments.
[0065] S2. The cells in the logarithmic growth phase were digested with trypsin and counted under a microscope to prepare 6×10 4 Take 100 μl of the cell suspension to 96-well culture plates, and inoculate two duplicate wells of each plate for each cell type as duplicate wells. 3 cells / well, 100 μl of culture medium was used as blank control, and the cells were cultured at 37°C overnight.
[0066] S3. Prepare a Co(DTC)3 complex stock solution with an initial concentration of 20 mM, dilute it according to a concentration gradient, and add three replicates of each concentration to a 96-well plate. After treating the cells for 72 hours, mix Cell Counting Kit-8 (CCK-8) and serum-free essential basic medium in a 1:10 volume ratio, add 100 μL per well to the wells to be tested, and incubate in a 37°C, 5% CO2 incubator for 1 hour.
[0067] S4. Absorbance at 450 nm was measured using a microplate reader, and the values for each plate were recorded. Cell viability in each group was determined using the CCK-8 assay, and the inhibition rate was calculated: inhibition rate = (OD value of the control group - OD value of the drug group) / (OD value of the control group) × 100%. Calculation of the half-maximal inhibitory concentration (IC50) values: GraphPad Prism 9 software was used to calculate the half-maximal inhibitory concentration (IC50) values based on the effects of different drug concentrations on cells, as shown in Table 1.
[0068] Table 1 IC50 of complexes (unit: μmol / L)
[0069]
[0070] From the data in Table 1, it can be seen that the disulfide metal ligand compounds Fe(DTC)3 and Co(DTC)3 can effectively inhibit the proliferation of three human tumor cells, namely lung cancer A459, liver cancer HEPG2, and gastric cancer SGC790 cells, with the IC50 values of 9.20, 12.16, and 8.06 μmol / L and 32.67, 26.84, and 18.42 μmol / L for 72 hours, respectively.
[0071] According to the synthesis method of the disulfide metal ligand compound Fe(DTC)3 in 1.1 Example 1, the corresponding ligand was used to replace sodium diethyldithiocarbamate to synthesize compounds Cu(CH2NS2)2, Cu(C3H5OS2)2, Cu(C7H5S2)2, Fe(CH2NS2)3, Fe(C3H5OS2)3, Co(CH2NS2)3, Co(C3H5OS2)3, Ni(DTC)3, Ni(CH2NS2)3, and Ni(C3H5OS2)3, respectively. The anti-tumor activity of compounds (H-1) to (H-10) was verified according to the experimental method of 1.2 Anti-tumor activity of the disulfide metal ligand compound Fe(DTC)3.
[0072] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by any technician familiar with this technical field within the technical scope disclosed by the present invention should be covered by the scope of protection of the present invention.
Claims
1. Use of a disulfide metal ligand compound in the preparation of an anti-tumor drug, wherein the chemical structure of the compound is shown in formula (I): in, R1 and R2 are each independently selected from any one of a hydrogen atom, an alkyl group, a substituted alkyl group, an alkenyl group, a substituted alkenyl group, an alkynyl group, a substituted alkynyl group, a monocyclic aromatic group, a substituted monocyclic aromatic group, a polycyclic aromatic group, a substituted polycyclic aromatic group, a polyheterocyclic aromatic group, a substituted polyheterocyclic aromatic group, and a polymer group containing a repeating unit structure of polyvinyl alcohol or polyethylene glycol and having a molecular weight of 100,000 g / mol or less; A is selected from C, N or O; M is a metal atom selected from any one of Fe, Co, Ni, Ru, Rh, Pd, Os, Ir, Pt, Mn, Tc, Re, Cr, Mo, W, V, Nb, Ta, Ti, Zr, Hf, Sc, Y, La, Ac, Cu, Zn, Ag, Cd, Au, Hg, Rg, Cn, Hs, Mt, Ds, Bh, Sg, Db, and Rf; n is an integer greater than or equal to 2.
2. The use according to claim 1, characterized in that R1 and R2 are each independently selected from hydrogen, C1-C4 alkyl, and monocyclic aromatic group; A is selected from N, O, and C; M is selected from Fe, Co, Ni, and Cu; and n is 2 or 3.
3. The use according to claim 2, characterized in that The chemical structure of the compound is shown in formula (II) or formula (III):
4. The use according to claim 2, characterized in that The chemical structure of the compound is selected from the following group:
5. The use according to claim 1, characterized in that The preparation method of the compound comprises the following steps: adding a metal salt solution dropwise into a disulfide ligand solution, reacting to generate a precipitate, and post-processing to obtain a disulfide metal ligand compound.
6. The use according to claim 5, characterized in that The molar ratio of the metal ions in the metal salt solution to the disulfide ligands in the disulfide ligand solution is n:
1.
7. The use according to claim 5, characterized in that Include one or more of the following characteristics: (a) stirring during the reaction; (b) the reaction temperature is room temperature; (c) the reaction time is more than 6 hours; (d) The post-treatment comprises filtering, washing and vacuum drying the precipitate.
8. The use according to claim 7, characterized in that Include one or more of the following characteristics: (d1) The filtration is performed by suction filtration using a Buchner funnel; (d2) the washing medium is selected from methanol, ethanol or ultrapure water; (d3) The number of washings is three times; (d4) The vacuum drying temperature is 40-60° C., and the vacuum drying time is 6-24 h.
9. The use according to claim 1, characterized in that The anti-tumor drug is a drug for lung cancer, liver cancer and gastric cancer.
10. The use according to claim 1, characterized in that The anti-tumor drug also includes a pharmaceutically acceptable carrier or excipient, or a combination thereof.